Evidence for the formation of a heterotrimeric complex of leukaemia inhibitory factor with its receptor subunits in solution
- Author(s)
- Zhang, JG; Owczarek, CM; Ward, LD; Howlett, GJ; Fabri, LJ; Roberts, BA; Nicola, NA;
- Journal Title
- BIOCHEMICAL JOURNAL
- Publication Type
- Journal Article
- Abstract
- Leukaemia inhibitory factor (LIF) is a polyfunctional cytokine that is known to require at least two distinct receptor components (LIF receptor alpha-chain and gp130) in order to form a high-affinity, functional, receptor complex. Human LIF binds with unusually high affinity to a naturally occurring mouse soluble LIF receptor alpha-chain, and this property was used to purify a stable complex of human LIF and mouse LIF receptor alpha-chain from pregnant-mouse serum. Recombinant soluble human gp130 was expressed, with a FLAG(R) epitope (DYKDDDDK) at the N-terminus, in the methylotropic yeast Pichia pastoris and purified using affinity chromatography. The formation of a trimeric complex in solution was established by native gel electrophoresis, gel-filtration chromatography, sedimentation equilibrium analysis, surface plasmon resonance spectroscopy and chemical crosslinking. The stoichiometry of this solution complex was 1:1:1, in contrast with that of the complex of interleukin-6, the interleukin-6-specific low-affinity receptor subunit and gp130, which is 2:2:2.
- Publisher
- PORTLAND PRESS
- Keywords
- IL-6 SIGNAL TRANSDUCER; ONCOSTATIN-M; HUMAN INTERLEUKIN-5; HEXAMERIC COMPLEX; LIF RECEPTOR; SOLUBLE FORM; MOUSE SERUM; GAMMA-CHAIN; GP130; BINDING
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Creation Date: 1997-08-01 12:00:00